Detection of protein-protein interactions on SiO2/Si surfaces by spectroscopic ellipsometry

Anal Biochem. 2003 Oct 1;321(1):65-70. doi: 10.1016/s0003-2697(03)00422-6.

Abstract

We have applied spectroscopic ellipsometry to sensitive detection of specific protein-protein interactions on SiO2/Si substrates. First, the change of ellipticity of the reflected polarized light (600-1100 nm) was correlated with the thickness of the protein layer immobilized on SiO2/Si surfaces by measuring monomeric (myoglobin) and homotetrameric (hemoglobin) proteins with a similar monomer size. Protein-protein interactions were then measured with the antigen/antibody and cell-surface receptor/ligand systems; in each system either of the two proteins was bound to SiO2/Si substrates. Consequently, significant ellipticity changes were observed only for the cases where the interactions were specific. A specific antibody binding was also detectable with an antigen displayed on the surface of bacteriophage particles. These results show the usefulness of spectroscopic ellipsometry for sensitive detection of protein-protein interactions and its applicability to a detection method for the protein-based biochips to be developed in the future.

MeSH terms

  • Animals
  • Antibodies / immunology
  • Antibody Specificity
  • Antigens / analysis
  • Antigens / immunology
  • Bacteriophage M13 / genetics
  • Epidermal Growth Factor / metabolism
  • ErbB Receptors / analysis
  • ErbB Receptors / metabolism
  • Hemoglobins / analysis
  • Hemoglobins / metabolism
  • Humans
  • Infrared Rays
  • Ligands
  • Mice
  • Myoglobin / analysis
  • Myoglobin / metabolism
  • Protein Binding
  • Proteins / analysis
  • Proteins / metabolism*
  • Rabbits
  • Recombinant Fusion Proteins / analysis
  • Recombinant Fusion Proteins / metabolism
  • Silicon / chemistry*
  • Silicon Dioxide / chemistry*
  • Spectrum Analysis / instrumentation*
  • Spectrum Analysis / methods*

Substances

  • Antibodies
  • Antigens
  • Hemoglobins
  • Ligands
  • Myoglobin
  • Proteins
  • Recombinant Fusion Proteins
  • Epidermal Growth Factor
  • Silicon Dioxide
  • ErbB Receptors
  • Silicon